甜瓜CmABCG8基因的表达特性分析

doi:10.13560/j.cnki.biotech.bull.1985.2021-1117

摘要/Abstract

摘要：

ABC转运蛋白,是一类功能广泛的具有主动介导多种分子转运的蛋白质,但在甜瓜（Cucumis melo L.）中的研究尚不清楚。本文通过生物信息学方法分析了ABC转运蛋白亚家族G成员CmABCG8的特点,同时使用荧光定量PCR检测了CmABCG8的不同时期果实、不同组织表达特性及对逆境胁迫的响应情况。进化树结果表明CmABCG8与黄瓜CsABCG23的亲缘关系最近,CmABCG8在甜瓜各组织中均有表达,在雌蕊和叶中表达量较高,CmABCG8有10个相互作用蛋白质。RT-qPCR结果表明,在果实不同时期的表达量中,CmABCG8在10 d时表达量最高;在甜瓜不同组织中,CmABCG8在叶子中表达量最高;分别用细胞分裂素、油菜素内酯、过氧化氢处理后,随着处理浓度的增加,CmABCG8的表达量均呈上升趋势;在铁、铜、锰3种金属离子处理后,CmABCG8的表达量呈下降趋势,因此分析其在甜瓜果实发育前期有一定影响,并对植物激素及金属离子有不同程度的响应,植物激素能够有效诱导CmABCG8的表达。

关键词: 甜瓜, ABC转运蛋白, 植物激素, 金属离子

Abstract:

ABC transporter is a kind of proteins with extensive functions,which can actively mediate the transport of various molecules,but its study in melon（Cucumis melo L.）is still unclear. In this paper,the characteristics of ABC transporter subfamily G member CmABCG8 were analyzed by bioinformatics method,and the expression characteristics of CmABCG8 in different fruits and tissues at different periods and their responses to stress were detected by fluorescence quantitative PCR. The phylogenetic tree results showed that CmABCG8 had the closest relationship with cucumber（Cucumis sativus L.）CsABCG23. CmABCG8 was expressed in all tissues of melon,and was highly expressed in the gynoecium and leaves. CmABCG8 had 10 interacting proteins. The results of RT-qPCR showed that the expression of CmABCG8 was the highest at 10 d. CmABCG8 expression was the highest in the leaves among different tissues of melon. After treatment with cytokinin,brassinosteroid and hydrogen peroxide,the expression of CmABCG8 presented an upward trend with the increase of the treatment concentration. After treatment with Fe²⁺,Cu²⁺ and Mn²⁺,the expression of CmABCG8 showed a downward trend. Therefore,it was analyzed that CmABCG8 had a certain influence on the early development of melon fruit and had different degrees of response to plant hormones and metal ions. Plant hormones could effectively induce the expression of CmABCG8.

Key words: Cucumis melo L., ABC transporter, plant hormone, metal ion

洪天澍, 海英, 恩和巴雅尔, 高峰. 甜瓜CmABCG8基因的表达特性分析[J]. 生物技术通报, 2022, 38(7): 178-185.

HONG Tian-shu, HAI Ying, ENHE Ba-ya-er, GAO Feng. Analysis of Expression Characteristics of CmABCG8 Gene in Cucumis melo L.[J]. Biotechnology Bulletin, 2022, 38(7): 178-185.

图/表 7

图1 CmABCG8和拟南芥、番茄、黄瓜、甜瓜中ABCG基因的系统进化树

Fig.1 Phylogenetic trees of ABCG genes in CmABCG8 and Arabidopsis thaliana,tomato（Solanum lycopersic-um）,cucumber（Cucumis sativus L.）and melon（Cucumis melo L.）

图2 甜瓜ABCG基因家族在不同组织中的表达

Fig. 2 Expression of ABCG gene family in the different tissues of melon

图3 甜瓜CmABCG8的蛋白质互作网络

Fig.3 Protein interaction network of CmABCG8 in melon

图4 甜瓜果实不同发育时期CmABCG8基因的相对表达量

Fig.4 Relative expressions of CmABCG8 gene in melon fruit at different development stages

图5 甜瓜不同组织中CmABCG8基因的相对表达量图中误差线表示标准偏差;星号代表显著差异（*P<0.05,** P <0.01）;下同

Fig.5 Relative expressions of CmABCG8 gene in the different tissues of melon The error line in the figure refers to the standard deviation;and * indicates significant difference（*P<0.05 and ** P <0.01）. The same below

图6 植物激素和H2O2处理对CmABCG8基因表达特性的影响 A：细胞分裂素处理;B：油菜素内酯处理;C：过氧化氢处理

Fig.6 Effects of plant hormones and H2O2 treatment on the expression characteristics of CmABCG8 gene A：Cytokinin treatment. B：Brassinosteroid treatment. C：Hydrogen peroxide treatment

图7 不同金属离子处理对CmABCG8基因表达特性的影响 A：Fe2+处理;B：Cu2+处理;C：Mn2+处理

Fig.7 Effects of different metal ion treatment on the expression characteristics of CmABCG8 gene A：Fe2+ treatment. B：Cu2+ treatment. C：Mn2+ treatment

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